LOCUS Exported 5930 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION E.Coli produced SAP25 SID purified by FPLC nickel column. ACCESSION . VERSION . KEYWORDS GB1-His-SAP25-126-186(pL91) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5930) AUTHORS Chandru A, Bate N, Vuister GW, Cowley SM TITLE Sin3A recruits Tet1 to the PAH1 domain via a highly conserved Sin3-Interaction Domain. JOURNAL Sci Rep. 2018 Oct 2;8(1):14689. doi: 10.1038/s41598-018-32942-w. PUBMED 30279502 REFERENCE 2 (bases 1 to 5930) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5930 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 20..44 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." RBS 59..81 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 95..256 /codon_start=1 /product="B1 domain of Streptococcal protein G " /label=GB1 /note="effective as a solubilizing fusion partner (Cheng and Patel, 2004)" /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD ATKTFTVTE" CDS 263..280 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 287..307 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 536..553 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 719..805 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 897..924 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" primer_bind complement(978..996) /label=T7 Term /note="T7 terminator, reverse primer" terminator 992..1039 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1076..1531 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1163..1182) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1373..1394 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(1646..2506) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 2269..2288 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2507..2599) /gene="bla" /label=AmpR promoter rep_origin 2680..3268 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3169..3188 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3422..3439 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3454..3593 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3519..3538) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3679..3701 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3695..3886) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4458..5540) /codon_start=1 /gene="lacI" /product="lac repressor" /label=lacI /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind 5502..5521 /label=LacI-R /note="LacI, reverse primer" promoter complement(5541..5618) /gene="lacI" /label=lacI promoter primer_bind 5824..5843 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt 61 tgtttaactt taagaaggag atatacatat gcagtacaaa ctgatcctga acggtaaaac 121 cctgaaaggt gaaaccacca ccgaagctgt tgacgctgct accgcggaaa aagttttcaa 181 acagtacgct aacgacaacg gtgttgacgg tgaatggacc tacgacgacg ctaccaaaac 241 cttcaccgtt accgaaggat ctcatcatca tcatcatcat ggatccgaaa acctgtattt 301 tcagggcgcc tcctccacct ggctcagcga ggcagagatg atcgcccttg ctggtctgct 361 gcagatgagc cagggagagc agacacccaa ctgcgtggca agctctctgc cttccaccag 421 ctgcccagac cctgtctctg tctctgaaga cccaggtccc agtggtgacc agagctgttc 481 tgggactgac acatgaaatt cgagctccgt cgacaagctt gcggccgcac tcgagcacca 541 ccaccaccac cactaatgtt aattaagttg ggcgttccta ggctgataaa acagaatttg 601 cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc 661 cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca 721 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt 781 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg 841 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa 901 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa 961 tacattcaaa tatgtatccg ctgagcaata actagcataa ccccttgggg cctctaaacg 1021 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg 1081 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 1141 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 1201 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 1261 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 1321 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 1381 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 1441 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 1501 aattttaaca aaatattaac gtttacaatt tctggcggca cgatggcatg agattatcaa 1561 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 1621 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 1681 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 1741 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 1801 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 1861 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 1921 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 1981 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 2041 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 2101 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 2161 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 2221 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 2281 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 2341 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 2401 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 2461 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 2521 aatcatgatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 2581 atttagaaaa ataaacaaat aggtcatgac caaaatccct taacgtgagt tttcgttcca 2641 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 2701 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 2761 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 2821 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 2881 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 2941 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 3001 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 3061 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 3121 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 3181 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 3241 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 3301 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 3361 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 3421 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 3481 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca 3541 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac 3601 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca 3661 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga 3721 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt 3781 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc 3841 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt 3901 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga 3961 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg 4021 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta 4081 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca 4141 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga 4201 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct 4261 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc 4321 tcaacgacag gagcacgatc atgctagtca tgccccgcgc ccaccggaag gagctgactg 4381 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta 4441 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 4501 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt 4561 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag 4621 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg 4681 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg 4741 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga 4801 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt 4861 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg 4921 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg 4981 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc 5041 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga 5101 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc 5161 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta 5221 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg 5281 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg 5341 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg 5401 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg 5461 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg 5521 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat 5581 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt 5641 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc 5701 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc 5761 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc 5821 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc 5881 cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat //